Effects of freezing and activation on membrane quality and DNA damage in Xenopus tropicalis and Xenopus laevis spermatozoa
S. Morrow A , J. Gosálvez B , C. López-Fernández B , F. Arroyo B , W. V. Holt C and M. J. Guille A D
+ Author Affiliations
- Author Affiliations
A School of Biological Sciences and European Xenopus Resource Centre, The University of Portsmouth, Portsmouth, PO1 2DT, UK.
B Department of Biology, Genetics Unit, The Autonomous University of Madrid, 20849 Madrid, Spain.
C Academic Department of Reproductive and Developmental Medicine, The University of Sheffield, Sheffield, S10 2SF, UK.
D Corresponding author. Email: matthew.guille@port.ac.uk
Reproduction, Fertility and Development 29(8) 1556-1566 https://doi.org/10.1071/RD16190
Submitted: 10 April 2015 Accepted: 4 July 2016 Published: 3 October 2016
Journal Compilation © Published Open Access CC BY 2017 Open Access CC BY-NC-ND
Abstract
There is growing concern over the effect of sperm cryopreservation on DNA integrity and the subsequent development of offspring generated from this cryopreserved material. In the present study, membrane integrity and DNA stability of Xenopus laevis and Xenopus tropicalis spermatozoa were evaluated in response to cryopreservation with or without activation, a process that happens upon exposure to water to spermatozoa of some aquatic species. A dye exclusion assay revealed that sperm plasma membrane integrity in both species decreased after freezing, more so for X. laevis than X. tropicalis spermatozoa. The sperm chromatin dispersion (SCD) test showed that for both X. tropicalis and X. laevis, activated frozen spermatozoa produced the highest levels of DNA fragmentation compared with all fresh samples and frozen non-activated samples (P < 0.05). Understanding the nature of DNA and membrane damage that occurs in cryopreserved spermatozoa from Xenopus species represents the first step in exploiting these powerful model organisms to understand the developmental consequences of fertilising with cryopreservation-damaged spermatozoa.
Additional keywords: cryopreservation, DNA fragmentation, sperm chromatin dispersion.
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